Nutritional Compositions Containing Punicalagins

ABSTRACT

The present invention relates generally to a nutritional composition comprising punicalagins.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates generally to nutritional compositionscomprising punicalagins and methods of using nutritional compositionscomprising punicalagins.

SUMMARY OF THE INVENTION

In an embodiment, the present invention is directed to nutritionalcompositions comprising a protein source, a fat source, a carbohydratesource, and punicalagins.

In another embodiment, the invention is directed to a method forenhancing the immune system in a pediatric subject via theadministration of punicalagins.

Further, the invention, in an embodiment, is directed to a method forreducing allergic inflammatory responses in a pediatric subject viaadministration of punicalagins.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

Reference now will be made in detail to the embodiments of theinvention, one or more examples of which are set forth below. Eachexample is provided by way of explanation of the invention, not alimitation of the invention. In fact, it will be apparent to thoseskilled in the art that various modifications and variations can be madein the present invention without departing from the scope or spirit ofthe invention. For instance, features illustrated or described as partof one embodiment, can be used on another embodiment to yield a stillfurther embodiment.

Thus, it is intended that the present invention covers suchmodifications and variations as come within the scope of the appendedclaims and their equivalents. Other objects, features and aspects of thepresent invention are disclosed in or are obvious from the followingdetailed description. It is to be understood by one of ordinary skill inthe art that the present discussion is a description of exemplaryembodiments only, and is not intended as limiting the broader aspects ofthe present invention.

The present invention is directed, in some embodiments, to nutritionalcompositions comprising a protein source, a fat source, a carbohydratesource, and punicalagins. The nutritional composition may be provided inany form known in the art, such as a powder, a gel, a suspension, apaste, a solid, a liquid, a liquid concentrate, or a ready-to-useproduct. The nutritional composition may, in certain embodiments,comprise a nutritional supplement, children's nutritional product,infant formula, human milk fortifier, or any other nutritionalcomposition designed for children or infants. As used herein, the terms“children” or “child” refer to human subjects between the ages of oneand 13 years old. In some embodiments, the terms “children” or “child”refer to human subjects that are two, three, four, five, or six yearsold. The term “children's nutritional product” means a composition thatsatisfies at least a portion of the nutrient requirements of a child. Asused herein, the term “infant” means a postnatal human that is less thanabout one year of age. The term “infant formula” means a compositionthat satisfies the nutrient requirements of an infant by being asubstitute for human milk.

Punicalagins are tannins, which are large polyphenol compounds that areisomers of 2,3-(S)-hexahydroxydiphenoyl-4,6-(S,S)-gallagyl-D-glucose,hydrolysable tannins with a molecular weight of 1084. Punicalagins arethe predominant pomegranate tannin.

In some embodiments of the invention, punicalagins are present in anutritional composition in an amount ranging from about 0.004 to about0.20 mg/g of the composition. In other embodiments of the invention,punicalagins are present in an amount ranging from about 0.008 to about0.12 mg/g of the composition.

If the composition of the invention is administered to an infant orchild, an amount of punicalagins ranging from about 1 mg to about 100 mgper day may be administered. In other embodiments, the amount ofpunicalagins administered to an infant or child via the composition ofthe invention may range from about 5 mg to about 50 mg per day. In someembodiments the amount of punicalagins administered to an infant orchild via the composition of the invention may range from about 10 mg toabout 35 mg per day.

In some embodiments, the nutritional composition of the presentinvention further comprises punicic acid. Punicic acid is apolyunsaturated fatty acid, 18:3 (n-5). It is obtained from pomegranateseed oil. Its chemical formula is C₁₈H₃₀O₂. Punicic acid is a conjugatedlinolenic acid. It has three conjugated double bonds, and it ischemically similar to the conjugated linoleic acids, which have two.

The term “punicic acid,” as used herein, refers to a conjugatedlinolenic acid isomer containing cis-9, trans-11, cis-13 double bonds inthe C₁₈ carbon chain, its non-toxic salts, active esters, activeisomers, active metabolites, structural lipids containing punicic acid,and mixtures thereof.

In some embodiments of the invention, punicic acid is present in anamount ranging from about 0.01 to about 40 mg/g of the composition. Infurther embodiments of the invention, punicic acid is present in anamount ranging from about 0.02 to about 20 mg/g of the composition.

If the composition of the invention is administered to an infant orchild, an amount of punicic acid ranging from about 25 mg to about 10 gper day may be administered. In further embodiments, the amount ofpunicic acid administered to an infant or child via the composition ofthe invention may range from about 50 mg to about 5 g per day.

If punicic acid is added to the nutritional composition of the presentinvention, the weight ratio of punicalagins:punicic acid may be fromabout 1:10,000 to about 4:1. In some embodiments of the presentinvention, the weight ratio of punicalagins:punicic acid may be fromabout 1:1,000 to about 1:1. In further embodiments, the ratio may befrom about 1:500 to about 2:3.

In additional embodiments of the invention, the nutritional compositionmay further comprise pomegranate leaf extract. Although previousresearch has focused on the pomegranate fruit, the extract frompomegranate leaves may also provide health benefits. The use ofpomegranate leaf extract in combination with punicalagins in accordancewith the present invention may provide increased levels of healthbenefits in infants, children and adult mammal subjects.

In some embodiments of the invention, pomegranate leaf extract ispresent in an amount ranging from about 4 mg/g to about 60 mg/g of thecomposition. In further embodiments of the invention, pomegranate leafextract is present in an amount ranging from about 12 mg/g to about 48mg/g of the composition.

If the composition of the invention is administered to an infant orchild, an amount of pomegranate leaf extract ranging from about 1 g toabout 15 g per day may be administered. In further embodiments, theamount of pomegranate leaf extract administered to an infant or childvia the composition of the invention may range from about 3 g to about12 g per day. In some embodiments, the amount of pomegranate leafextract administered to an infant or child via the composition of theinvention may range from about 5 g to about 10 g per day.

If the nutritional composition contains pomegranate leaf extract, theweight ratio of punicalagins:pomegranate leaf extract may be from about1:15,000 to about 1:100. In some embodiments of the present invention,the weight ratio of punicalagins:pomegranate leaf extract may be fromabout 1:2,400 to about 1:60. In further embodiments, the ratio may befrom about 1:500 to about 1:10.

In further embodiments, the nutritional composition of the presentinvention contemplates the use of punicalagins with punicic acid andpomegranate leaf extract. The combination of punicalagins, punicic acid,and pomegranate leaf extract may provide increased levels of healthbenefits in infants, children, and adult mammal subjects.

If punicalagins are provided in an infant formula or children'snutritional product, the formula or product may be nutritionallycomplete and may contain suitable types and amounts of lipid,carbohydrate, protein, vitamins, and minerals. As used herein, the term“nutritionally complete” refers to a nutritional composition that may beused as the sole source of nutrition, which would supply to a subjectessentially all the required daily amounts of vitamins, minerals, and/ortrace elements in combination with the proteins, carbohydrates, andlipids.

If the composition of the present invention is provided as an infantformula, the amount of lipid or fat in an infant formula can vary fromabout 3 to about 7 g/100 kcal. Lipid sources can be any used in the art,e.g., vegetable oils such as palm oil, canola oil, corn oil, soybeanoil, palmolein, coconut oil, medium chain triglyceride oil, high oleicsunflower oil, high oleic safflower oil, fish oil, and the like. Theamount of carbohydrate can vary from about 8 to about 12 g/100 kcal.Carbohydrate sources can be any used in the art, e.g., lactose, glucose,corn syrup solids, maltodextrins, sucrose, starch, rice syrup solids,and the like.

The amount of protein, if the present composition is provided as aninfant formula, can vary from about 1 to about 5 g/100 kcal. Proteinsources can be any used in the art, e.g., nonfat milk, whey protein,casein, soy protein, hydrolyzed protein, amino acids, and the like. In aparticular embodiment, the protein source contains both bovine whey andbovine casein. In an embodiment, the whey may be a denatured wheyprotein concentrate. In another embodiment, the whey may be anundenatured whey protein concentrate. In a specific embodiment, theprotein source may contain both a denatured and an undenatured wheyprotein concentrate.

If the composition of the present invention is provided as an infantformula, suitable examples of infant formulas may include soy based,partially or extensively hydrolyzed, lactose-free, low-lactose,elemental, protein-free, anti-regurgitation, premature, or standardinfant formulas. The type of infant formula used may be based on theneeds of the infant for proper digestion and health.

The composition of the present invention may be provided to both termand preterm infants. As a result, if the composition of the presentinvention is provided as an infant formula, the infant formula may be aterm infant formula or a preterm infant formula. Similarly, if thecomposition of the present invention is provided as a human milkfortifier, the human milk fortifier may be a term human milk fortifieror a preterm human milk fortifier.

Long chain polyunsaturated fatty acids (LCPUFA) have been shown to beimportant in infant development. Arachidonic acid (ARA; C20:4, n-6) anddocosahexaenoic acid (DHA; C22:6 n-3) are of particular interest due tothe high concentrations of each found in the infant brain and retina.ARA and DHA are synthesized from their respective 18 carbon precursors,linoleic acid (18:2, n-6) and a-linolenic acid (18:3, n-3) throughalternate desaturation and elongation. DHA and ARA are typicallyobtained through breast milk in infants that are breast-fed. In infantsthat are formula-fed, however, DHA and ARA must be supplemented into thediet.

Evidence indicates that infants with altered LCPUFA levels, resultingfrom inadequate intake of dietary LCPUFA, may be at risk forneurological problems, may score lower on cognitive tests, and havelower retinal development than infants fed human milk. Thus, theprovision of LCPUFA, in particular ARA and DHA, in amounts closelyapproximating those found in human milk may support adequate growth aswell as neurological development in formula-fed infants. Thus, in someembodiments of the invention, the nutritional composition contains atleast one LCPUFA. In a particular embodiment, the nutritionalcomposition contains DHA and/or ARA.

If added to a nutritional composition, the weight ratio of ARA:DHA maybe from about 1:3 to about 9:1. In one embodiment of the presentinvention, this ratio is from about 1:2 to about 4:1. In yet anotherembodiment, the ratio is from about 2:3 to about 2:1. In one particularembodiment the ratio is about 2:1. In another particular embodiment ofthe invention, the ratio is about 1:1.5. In other embodiments, the ratiois about 1:1.3. In other embodiments, the ratio is about 1:1.9. In aparticular embodiment, the ratio is about 1.5:1. In a furtherembodiment, the ratio is about 1.47:1.

If DHA is included in the invention, the level of DHA may be betweenabout 0.0% and 1.00% of fatty acids, by weight. In other embodiments,the level of DHA may be about 0.32% by weight. In some embodiments, thelevel of DHA may be about 0.33% by weight. In another embodiment, thelevel of DHA may be about 0.64% by weight. In another embodiment, thelevel of DHA may be about 0.67% by weight. In yet another embodiment,the level of DHA may be about 0.96% by weight. In a further embodiment,the level of DHA may be about 1.00% by weight.

If ARA is included in the invention, the level of ARA may be between0.0% and 0.67% of fatty acids, by weight. In another embodiment, thelevel of ARA may be about 0.67% by weight. In another embodiment, thelevel of ARA may be about 0.5% by weight. In yet another embodiment, thelevel of DHA may be between about 0.47% and 0.48% by weight.

If used, the amount of DHA in the present invention may be from about 2mg/100 kilocalories (kcal) to about 100 mg/100 kcal. In anotherembodiment, the amount of DHA may be from about 5 mg/100 kcal to about75 mg/100 kcal. In yet another embodiment, the amount of DHA may be fromabout 15 mg/100 kcal to about 60 mg/100 kcal.

If used, the amount of ARA in the present invention may be from about 4mg/100 kilocalories (kcal) to about 100 mg/100 kcal. In anotherembodiment, the amount of ARA may be from about 10 mg/100 kcal to about67 mg/100 kcal. In yet another embodiment, the amount of ARA may be fromabout 20 mg/100 kcal to about 50 mg/100 kcal. In a particularembodiment, the amount of ARA may be from about 25 mg/100 kcal to about40 mg/100 kcal. In one embodiment, the amount of ARA is about 30 mg/100kcal.

If the composition of the invention is supplemented with oils containingDHA and/or ARA, it may be accomplished using standard techniques knownin the art. For example, an equivalent amount of an oil which isnormally present in the composition may be replaced with DHA and/or ARA.

If utilized, the source of one or more of the LCPUFA can be any sourceknown in the art such as marine oil, fish oil, single cell oil, egg yolklipid, brain lipid, and the like. Any LCPUFA can be in natural form,provided that the remainder of the LCPUFA source does not result in anysubstantial deleterious effect on an infant or child. Alternatively, aLCPUFA can be used in refined form.

If used, the LCPUFA source may or may not contain eicosapentaenoic acid(EPA). In some embodiments, especially for feeding to infants the LCPUFAused in the invention contains little or no EPA. For example, in certainembodiments that the nutritional compositions used herein contain lessthan about 20 mg/100 kcal EPA; in some embodiments less than about 10mg/100 kcal EPA; in other embodiments less than about 5 mg/100 kcal EPA;and in still other embodiments substantially no EPA.

In some embodiments of the invention, the nutritional compositioncontains additional components which may include probiotics orprebiotics. The term “probiotic” means a microorganism that exertsbeneficial effects on the health of the host. Any probiotic known in theart may be added, provided it is suitable for combination with the othercomponents of the composition. For example, the probiotic may be chosenfrom the group consisting of Lactobacillus and Bifidobacterium.Alternatively, the probiotic can be Lactobacillus rhamnosus GG.

In certain embodiments, the nutritional composition of the presentinvention additionally comprises at least one prebiotic. The term“prebiotic”, as used herein, means a non-digestible food ingredient thatstimulates the growth and/or activity of probiotics. In this embodiment,any prebiotic known in the art may be added, provided it is suitable forcombination with the other components of the composition. In aparticular embodiment, the prebiotic can be selected from the groupconsisting of fructo-oligosaccharide, inulin, gluco-oligosaccharide,galacto-oligosaccharide, isomalto-oligosaccharide, xylo-oligosaccharide,soybean oligosaccharides, chito-oligosaccharide, gentio-oligosaccharide,manno-oligosaccharide, lactulose, lactosucrose, raffinose,aribino-oligosaccharide, glucans, siallyl-oligosaccharide, polydextrose,inulin, fuco-oligosaccharide, and mixtures thereof.

In some embodiments, the use of punicalagins alone, or in combinationwith punicic acid or pomegranate leaf extract, or the combination of allthree aid in immune system development and function in pediatricsubjects. For example, the use of punicalagins alone, or in combinationwith punicic acid or pomegranate leaf extract, or the combination of allthree may enhance resistance to infection and/or reduce allergicinflammatory responses including, but not limited to asthma, wheezing,atopic cough, bronchiolitis, bronchitis, and eczema. Thus, in someembodiments, the invention is directed to a method for enhancing theimmune response in a pediatric subject comprising administering thenutritional composition of the present invention to the pediatricsubject. In further embodiments, the invention is directed to a methodfor enhancing resistance to infection in a pediatric subject comprisingadministering the nutritional composition of the present invention tothe pediatric subject. In other embodiments, the invention is directedto a method for reducing allergic inflammatory responses in a pediatricsubject comprising administering the nutritional composition of thepresent invention to the pediatric subject. As used herein, the term“pediatric subject” refers to human subjects that are less than 13 yearsold. In some embodiments, the term “pediatric subject” refers to humansubjects that are less than 8 years old.

Further, in some embodiments, the use of punicalagins alone or incombination with punicic acid, or pomegranate leaf extract, or thecombination of all three may aid in enhancing cardiovascular health, eyehealth, brain development and function, gastrointestinal health andfunction. Further, the use of punicalagins alone, or in combination withpunicic acid or pomegranate leaf extract, or the combination of allthree may aid in reducing the risk of inflammation, cancer, or themetabolic syndrome, including obesity and diabetes mellitus. Thus, insome embodiments, the invention is directed to a method for enhancingcardiovascular health, and eye health in a pediatric subject byadministering the nutritional composition of the present invention tothe pediatric subject. In further embodiments the invention is directedto a method for enhancing brain development and function in a pediatricsubject by administering the nutritional composition of the presentinvention to the pediatric subject. In other embodiments the inventionis directed to a method of improving gastrointestinal health andfunction in a pediatric subject by administering the nutritionalcomposition of the present invention to the pediatric subject. Such anembodiment of the invention could include providing a healthy intestinalmicro flora balance in the pediatric subject. In some embodiments, theinvention is directed to a method for reducing the risk of inflammationand cancer in a pediatric subject by administering the nutritionalcomposition of the present invention to the pediatric subject. Infurther embodiments, the invention is directed to a method for reducingthe risk of metabolic syndrome in a pediatric subject by administeringthe nutritional composition of the present invention to the pediatricsubject. In such an embodiment, the reduction of the risk of metabolicsyndrome in pediatric subjects could include reduction in the risk ofobesity and diabetes mellitus.

In certain embodiments, the combination of punicalagins with punicicacid or with pomegranate leaf extract, or the combination of all threeprovides an enhanced effect with respect to the health benefits listedabove. It is believed that the activity of the combination ofpunicalagins with punicic acid or with pomegranate leaf extract, or thecombination of all three is greater than the added activity expectedwhen each of the compounds are administered separately.

The following example describes an embodiment of the present invention.Other embodiments within the scope of the claims herein will be apparentto one skilled in the art from the consideration of the specification orpractice of the invention disclosed herein. It is intended that thespecification, together with the example, be considered to be exemplaryonly, with the scope of the invention being indicated by the claimswhich follow the example.

EXAMPLE 1

Table 1 illustrates the nutrient components of one infant formulaembodiment of a nutritional composition of the present invention.

TABLE 1 Per 100 Calories (Normal Dilution) (5 fl oz) Per 100 gramsPowder Protein, g 2.1 10.8 Fat, g 5.3 27 Linoleic acid, mg 860 4400Linolenic acid, mg 85 440 DHA, mg 17 88 ARA, mg 34 175 Carbohydrate, g10.9 56 Water, g 134 3.3 Punicalagins, mg 1.9 10 Vitamins/OtherNutrients Vitamin A, IU 300 1550 Vitamin D, IU 60 310 Vitamin E, IU 210.3 Vitamin K, mcg 8 41 Thiamin (Vitamin B1), mcg 80 410 Riboflavin(Vitamin B2), 140 720 mcg Vitamin B6, mcg 60 310 Vitamin B12, mcg 0.31.55 Niacin, mcg 1000 5200 Folic acid (Folacin), mcg 16 83 Pantothenicacid, mcg 500 2600 Biotin, mcg 3 15.5 Vitamin C (Ascorbic acid), 12 62mg Choline, mg 24 124 Inositol, mg 6 31 Carnitine, mg 2 10.3 Taurine, mg6 31 Minerals Calcium, mg 78 400 Phosphorus, mg 43 220 Magnesium, mg 841 Iron, mg 1.8 9.3 Zinc, mg 1 5.2 Manganese, mcg 15 77 Copper, mcg 75390 Iodine, mcg 10 52 Selenium, mcg 2.8 14.5 Sodium, mg 27 139Potassium, mg 108 560 Chloride, mg 63 330 Molybdenum, mcg NA NAChromium, mcg NA NA

All references cited in this specification, including withoutlimitation, all papers, publications, patents, patent applications,presentations, texts, reports, manuscripts, brochures, books, internetpostings, journal articles, periodicals, and the like, are herebyincorporated by reference into this specification in their entireties.The discussion of the references herein is intended merely to summarizethe assertions made by their authors and no admission is made that anyreference constitutes prior art. Applicants reserve the right tochallenge the accuracy and pertinence of the cited references.

These and other modifications and variations to the present inventionmay be practiced by those of ordinary skill in the art, withoutdeparting from the scope of the present invention, which is moreparticularly set forth in the appended claims. In addition, it should beunderstood that aspects of the various embodiments may be interchangedin whole or in part. Furthermore, those of ordinary skill in the artwill appreciate that the foregoing description is by way of exampleonly, and is not intended to limit the invention so further described insuch appended claims. Therefore, the scope of the appended claims shouldnot be limited to the description of the preferred versions containedtherein.

1. A nutritional composition comprising: a. a protein source; b. a fatsource; c. a carbohydrate source; and d. punicalagins.
 2. Thenutritional composition according to claim 1 wherein the amount ofpunicalagins in the composition is between about 0.004 to about 0.20mg/g of the composition.
 3. The nutritional composition according toclaim 1, further comprising punicic acid.
 4. The nutritional compositionaccording to claim 3, wherein the amount of punicic acid in thecomposition is between about 0.01 to about 40 mg/g of the composition.5. The nutritional composition according to claim 1, further comprisingpomegranate leaf extract.
 6. The nutritional composition according toclaim 5, wherein the amount of pomegranate leaf extract in thecomposition is between about 4 mg/g to about 60 mg/g of the composition.7. The nutritional composition according to claim 1, further comprisingpunicic acid and pomegranate leaf extract.
 8. The nutritionalcomposition according to claim 1, wherein the nutritional composition isan infant formula.
 9. A method of enhancing the immune system of apediatric subject comprising administering punicalagins to the pediatricsubject.
 10. The method of claim 9, wherein the enhanced immune systemfunction comprises an improved resistance to infection.
 11. The methodaccording to claim 9, wherein the amount of punicalagins administered tothe pediatric subject is between about 1 mg to about 100 mg per day. 12.The method according to claim 9, wherein the method further comprisesadministering punicic acid to the pediatric subject.
 13. The methodaccording to claim 12, wherein the amount of punicic acid administeredto the pediatric subject is between about 50 mg to about 5 g per day.14. The method according to claim 9, wherein the method furthercomprises administering pomegranate leaf extract to the pediatricsubject.
 15. The method according to claim 14, wherein the amount ofpomegranate leaf extract administered to the pediatric subject isbetween about 1 g to about 15 g per day.
 16. The method according toclaim 9, wherein the method further comprises administering punicic acidand pomegranate leaf extract to the pediatric subject.
 17. A method forreducing allergic inflammatory responses in a pediatric subjectcomprising administering punicalagins to the pediatric subject.
 18. Themethod according to claim 17, wherein the amount of punicalaginsadministered to the pediatric subject is between about 1 mg to about 100mg per day.
 19. The method according to claim 17, wherein the methodfurther comprises administering punicic acid to the pediatric subject.20. The method according to claim 17, wherein the method furthercomprises administering pomegranate leaf extract to the pediatricsubject.